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Brevibacillus brevis H3 and application thereof

A technology of Bacillus brevis and decomposing bacteria agent, applied in the direction of application, bacteria, biochemical equipment and methods, etc., can solve the problems of low compost fermentation efficiency, achieve high protease activity, less environmental pollution, and rapid growth

Inactive Publication Date: 2015-03-04
HANGZHOU FAMOXI BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the composting process is greatly affected by factors such as microbial species and ambient temperature, the composting fermentation efficiency is low without the artificial addition of bacterial agents

Method used

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  • Brevibacillus brevis H3 and application thereof
  • Brevibacillus brevis H3 and application thereof
  • Brevibacillus brevis H3 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 strain screening and identification

[0057] 1. Screening and identification of strain H4

[0058] (1) Culture medium and reagent formula:

[0059] The final concentration of SOC liquid medium is composed of: tryptone 20g / L, yeast extract 5g / L, NaCl 0.5g / L, KCl 2.5mmol / L, MgCl 2 0.01mol / L, glucose 0.02mol / L, solvent is deionized water, pH value 7.0.

[0060] The final concentration of the SOC solid medium is composed of: SOC liquid medium plus agar powder with a final mass concentration of 1.5%.

[0061] Preparation method of SOC liquid medium: For each liter of medium, add 20g of tryptone, 5g of yeast extract, and 0.5g of NaCl into 965ml of deionized water and shake the container to completely dissolve the solute. Add 10ml 250mmol / L KCl aqueous solution, use 5mol / L NaOH to adjust the pH to 7.0, at 15psi (1.05kg / cm 2 ) steam sterilization under high pressure for 20min. Add 5ml filter-sterilized 2mol / L MgCl to the sterilized solution 2 , cooled to 60°C or...

Embodiment 2

[0153] Embodiment 2 decomposing bacteria agent

[0154] (1) Bacillus subtilis H4 freeze-dried powder

[0155] a. Incline cultivation

[0156] Bacillus subtilis H4 was inoculated into slant LB medium and cultured at 37°C for 1 day to obtain slant cells.

[0157] b. Seed cultivation

[0158] Pick an inoculation loop strain from the slant bacteria, inoculate it into 100ml LB liquid medium, culture it on a shaker at 50°C for 24 hours, and obtain a seed solution.

[0159] c. Fermentation culture

[0160] The seed solution was inoculated into the fermentation medium at a volume ratio of 1:1000, and fermented at 50°C for 24 hours. The pressure of the fermenter was 0.04MPa, the ventilation rate was 0.2v / v.min, and the rotation speed was 200rpm. The fermented liquid was taken and centrifuged, the wet cells were collected, and freeze-dried at -50°C for 24 hours to obtain the freeze-dried powder of Bacillus subtilis H4.

[0161] The final concentration of the fermentation medium is ...

Embodiment 3

[0170] The character of embodiment 3 decomposing bacterial agents

[0171] (1) Protease activity

[0172] Prepare L-tyrosine standard solutions of 10, 20, 30, 40, 50, and 60 μg / ml, use double-distilled water without tyrosine as a blank control, and measure the absorbance at a wavelength of 275 nm with an ultraviolet spectrophotometer. Taking the absorbance as the ordinate and the concentration of tyrosine as the abscissa, a standard curve was drawn to obtain the equation y=0.0074x, and the absorption constant K was 135.1.

[0173] Take 10 g of the decomposed microbial agent prepared by the method in Example 2, add 100 ml of 0.1 mol / L phosphate buffer solution (pH=7.5), and shake on a shaker at 200 rpm for 30 min. Centrifuge at 3000rpm and take the decomposing agent supernatant.

[0174] Take 1.0ml of decomposing agent supernatant, keep it warm at 40°C for 2 minutes, add 1.0ml casein aqueous solution (10mg / ml), mix well and react at 40°C for 20min, add 2ml trichloroacetic aci...

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Abstract

The invention discloses brevibacillus brevis H3 and application thereof to prepare a composting inoculant. The brevibacillus brevis H3 is preserved in China Center for Type Culture Collection with the preservation number of CCTCC NO: M2013481, the preservation data is 21rt, October, 2013, and the address is Wuhan University, Wuhan City, Hubei province. The brevibacillus brevis H3 is resistant to high temperature, is survival at a temperature scope of 30 DEG C-60 DEG C, and is relatively high in lipase activity and relatively rapid in growth. The composting inoculant prepared by utilizing the bacterial strain has the advantages of high temperature resistance and capability of secreting protease and lipase by efficiently fermenting animal wastes. By utilizing the composting inoculant with the advantages of being small in environmental pollution, economic, practicable and the like, after a product subjected to compost processing is subjected to subsequent processing, the processed product can be used as an organic fertilizer and the purpose of changing waste into valuables is reached.

Description

(1) Technical field [0001] The invention relates to a strain of Bacillus brevis and its application, in particular to a strain of Bacillus brevis H3 and its application in the preparation of decomposing bacterial agents. (2) Background technology [0002] Organic waste is often sent to landfills, incinerators or discarded as garbage. It is very necessary to develop methods for utilizing organic waste, not only to protect the environment, but also to turn waste into wealth. At present, many methods for utilizing organic waste have been developed, but most of them are for the utilization of waste with high cellulose and starch content, while there are fewer methods for the utilization of waste with high protein and fat content. [0003] Wastes rich in protein and fat (such as animal waste), if discarded directly, will generate a large amount of sewage and odor, pollute the environment, and may cause the spread of germs, posing a great safety hazard. At present, the methods f...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/00C12R1/01C12R1/125
CPCC05F11/08C12N1/205C12R2001/01Y02W30/40
Inventor 黄黎锋李海峰蓝袁洋施建军钟丹成沈如玥
Owner HANGZHOU FAMOXI BIOMEDICAL TECH
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